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. Author manuscript; available in PMC: 2014 May 14.
Published in final edited form as: Biochem J. 2014 Feb 15;458(1):95–107. doi: 10.1042/BJ20130685

Figure 7. MG-132 enhanced the interaction between hCAR1 and endogenous HSP90 in cytosolic fractions.

Figure 7

COS-1 cells were transfected with 3 × FLAG-hCAR1 as described in the Experimental section. After 36 h, the cells were treated with vehicle (0.1 % DMSO), PB (1 mM), 10 µM MG-132 (MG) alone, or co-treated with MG-132 plus PB. After 5 h treatment, cytosolic fractions were extracted and immunoprecipitated with anti-(HSP90 IgM) antibody plus Protein L-Agarose. The precipitated protein was subjected to Western blotting with anti-FLAG antibody, followed by anti-HSP90 antibody for endogenous HSP90 detection. The anti-HSP90 antibody precipitated equal amounts of HSP90 from the COS-1 cell cytosolic fractions, whereas control IgM did not precipitate HSP90. IP, immunoprecipitation; WB, Western blot.